Critical appraisal of four IL-6 immunoassays.

Thompson, Dana K

Huffman, Kim M

Kraus, William E

Kraus, Virginia Byers

Stoddart, Cheryl A

United States

2015-11-10T22:41:06Z

2012

BACKGROUND: Interleukin-6 (IL-6) contributes to numerous inflammatory, metabolic, and physiologic pathways of disease. We evaluated four IL-6 immunoassays in order to identify a reliable assay for studies of metabolic and physical function. Serial plasma samples from intravenous glucose tolerance tests (IVGTTs), with expected rises in IL-6 concentrations, were used to test the face validity of the various assays. METHODS AND FINDINGS: IVGTTs, administered to 14 subjects, were performed with a single infusion of glucose (0.3 g/kg body mass) at time zero, a single infusion of insulin (0.025 U/kg body mass) at 20 minutes, and frequent blood collection from time zero to 180 minutes for subsequent Il-6 measurement. The performance metrics of four IL-6 detection methods were compared: Meso Scale Discovery immunoassay (MSD), an Invitrogen Luminex bead-based multiplex panel (LX), an Invitrogen Ultrasensitive Luminex bead-based singleplex assay (ULX), and R&D High Sensitivity ELISA (R&D). IL-6 concentrations measured with MSD, R&D and ULX correlated with each other (Pearson Correlation Coefficients r = 0.47-0.94, p<0.0001) but only ULX correlated (r = 0.31, p = 0.0027) with Invitrogen Luminex. MSD, R&D, and ULX, but not LX, detected increases in IL-6 in response to glucose. All plasma samples were measurable by MSD, while 35%, 1%, and 4.3% of samples were out of range when measured by LX, ULX, and R&D, respectively. Based on representative data from the MSD assay, baseline plasma IL-6 (0.90 ± 0.48 pg/mL) increased significantly as expected by 90 minutes (1.29 ± 0.59 pg/mL, p = 0.049), and continued rising through 3 hours (4.25 ± 3.67 pg/mL, p = 0.0048). CONCLUSION: This study established the face validity of IL-6 measurement by MSD, R&D, and ULX but not LX, and the superiority of MSD with respect to dynamic range. Plasma IL-6 concentrations increase in response to glucose and insulin, consistent with both an early glucose-dependent response (detectable at 1-2 hours) and a late insulin-dependent response (detectable after 2 hours).

http://www.ncbi.nlm.nih.gov/pubmed/22347395

PONE-D-11-11488

1932-6203

https://hdl.handle.net/10161/10874

eng

Public Library of Science (PLoS)

PLoS One

10.1371/journal.pone.0030659

Glucose

Humans

Immunoassay

Insulin

Interleukin-6

Kinetics

Time Factors

Critical appraisal of four IL-6 immunoassays.

Journal article

Kraus, William E|0000-0003-1930-9684

Kraus, Virginia Byers|0000-0001-8173-8258

http://www.ncbi.nlm.nih.gov/pubmed/22347395

e30659

2

Biomedical Engineering

Clinical Science Departments

Duke

Duke Cancer Institute

Duke Molecular Physiology Institute

Institutes and Centers

Medicine

Medicine, Cardiology

Medicine, Rheumatology and Immunology

Orthopaedics

Pathology

Pratt School of Engineering

School of Medicine

School of Nursing

School of Nursing - Secondary Group

Published

7